Elizabeth M. Brown

Phylogenetic analysis reveals a cryptic species Blastomyces gilchristii, sp. nov. within the human pathogenic fungus Blastomyces dermatitidis.

Background Analysis of the population genetic structure of microbial species is of fundamental importance to many scientific disciplines because it can identify cryptic species, reveal reproductive mode, and elucidate processes that contribute to pathogen evolution. Here, we examined the population genetic structure and geographic differentiation of the sexual, dimorphic fungus Blastomyces dermatitidis, the causative agent of blastomycosis. Methodology/Principal Findings Criteria for Genealogical Concordance Phylogenetic Species Recognition (GCPSR) applied to seven nuclear loci (arf6, chs2, drk1, fads, pyrF, tub1, and its-2) from 78 clinical and environmental isolates identified two previously unrecognized phylogenetic species. Four of seven single gene phylogenies examined (chs2, drk1, pyrF, and its-2) supported the separation of Phylogenetic Species 1 (PS1) and Phylogenetic Species 2 (PS2) which were also well differentiated in the concatenated chs2-drk1-fads-pyrF-tub1-arf6-its2 genealogy with all isolates falling into one of two evolutionarily independent lineages. Phylogenetic species were genetically distinct with interspecific divergence 4-fold greater than intraspecific divergence and a high Fst value (0.772, P<0.001) indicative of restricted gene flow between PS1 and PS2. Whereas panmixia expected of a single freely recombining population was not observed, recombination was detected when PS1 and PS2 were assessed separately, suggesting reproductive isolation. Random mating among PS1 isolates, which were distributed across North America, was only detected after partitioning isolates into six geographic regions. The PS2 population, found predominantly in the hyper-endemic regions of northwestern Ontario, Wisconsin, and Minnesota, contained a substantial clonal component with random mating detected only among unique genotypes in the population. Conclusions/Significance These analyses provide evidence for a genetically divergent clade within Blastomyces dermatitidis, which we use to describe a novel species, Blastomyces gilchristii sp. nov. In addition, we discuss the value of population genetic and phylogenetic analyses as a foundation for disease surveillance, understanding pathogen evolution, and discerning phenotypic differences between phylogenetic species.

Rapid identification of herd effects with the introduction of serogroup C meningococcal conjugate vaccine in Ontario, Canada, 2000-2006.

In 2001, Canadas National Advisory Committee on Immunization endorsed a meningococcal serogroup C conjugate vaccine, which appears to provide durable serogroup-specific immunity while reducing nasopharyngeal carriage. With reference to direct and indirect effects on case occurrence, we sought to evaluate recent trends in the incidence of invasive meningococcal disease (IMD) in Ontario. Analyses included all IMD cases reported between 2000 and 2006 to the Ontario Central Public Health Laboratory. Poisson models incorporating terms for age, sex and seasonal oscillation identified a significant downward trend in disease occurrence, which was strongest in serogroup C cases and not evident when serogroup C strains were excluded from the analysis. Among age groups not targeted by the vaccine program serogroup C, IMD displayed a pattern of decreasing incidence that was not present in non-serogroup C disease. These apparent dramatic effects of conjugate C vaccine (both direct and indirect) may be important in the implementation and evaluation of vaccine policy in other jurisdictions.

Epidemiology of Invasive Meningococcal Disease with Decreased Susceptibility to Penicillin in Ontario, Canada, 2000 to 2006

ABSTRACT Neisseria meningitidis has been relatively slow to acquire resistance to penicillin. We previously reported an increase in the incidence of invasive meningococcal disease (IMD) strains with decreased susceptibility to penicillin (DSP) in Ontario. Our objectives were to evaluate trends in IMD with DSP, to identify case-level predictors of IMD with DSP, and to evaluate the relationship among DSP, bacterial phenotype, and the likelihood of a fatal outcome. All IMD isolates received in Ontario between 2000 and 2006 were submitted to the Public Health Laboratories, Toronto, for confirmation of the species, serogroup determination, and susceptibility testing. Isolates were considered to be IMD strains with DSP if the penicillin MIC was ≥0.125 μg/ml. Temporal trends were evaluated using multivariable Poisson regression models. Correlates of diminished susceptibility and fatal outcome were evaluated with multivariable logistic regression models. The overall rate of IMD caused by strains with DSP in Ontario was approximately 1.20 cases per million population annually (95% confidence interval [95% CI], 0.99 to 1.46). Seventy-nine strains (21.7%) were IMD strains with DSP. There was no year-to-year trend in the incidence of IMD with DSP. IMD with DSP was strongly associated with strains of serogroups Y (odds ratio [OR], 6.3; 95% CI, 3.6 to 11.1) and W-135 (OR, 8.2; 95% CI, 4.0 to 16.7). Infection with serogroup B or C strains was associated with a marked increase in the risk of mortality (OR, 3.07; 95% CI, 1.39 to 6.75); however, no association between IMD with DSP and mortality was observed. In contrast to trends of the 1990s, the incidence of IMD with DSP was stable in Ontario between 2000 and 2006. In Ontario, the serogroup rather than the penicillin MIC is the microbiological parameter most predictive of mortality.

Identification of sexual networks through molecular typing of quinolone-resistant Neisseria gonorrhoeae in Ontario, Canada.

Neisseria gonorrhoeae multi-antigen sequence typing technique demonstrated multiple sexual transmission networks in Ontario, Canada. Isolates with novel sequences had higher odds of originating in Toronto but had no association with heightened population-level quinolone exposure. Neisseria gonorrhoeae multi-antigen sequence typing technique can be a useful tool for investigation of multidrug-resistant N. gonorrhoeae emergence in North America.

Phylogeographic Analysis of Blastomyces dermatitidis and Blastomyces gilchristii Reveals an Association with North American Freshwater Drainage Basins

Blastomyces dermatitidis and Blastomyces gilchristii are dimorphic fungal pathogens that cause serious pulmonary and systemic infections in humans. Although their natural habitat is in the environment, little is known about their specific ecologic niche(s). Here, we analyzed 25 microsatellite loci from 169 strains collected from various regions throughout their known endemic range in North America, representing the largest and most geographically diverse collection of isolates studied to date. Genetic analysis of multilocus microsatellite data divided the strains into four populations of B. dermatitidis and four populations of B. gilchristii. B. dermatitidis isolates were recovered from areas throughout North America, while the B. gilchristii strains were restricted to Canada and some northern US states. Furthermore, the populations of both species were associated with major freshwater drainage basins. The four B. dermatitidis populations were partitioned among (1) the Nelson River drainage basin, (2) the St. Lawrence River and northeast Atlantic Ocean Seaboard drainage basins, (3) the Mississippi River System drainage basin, and (4) the Gulf of Mexico Seaboard and southeast Atlantic Ocean Seaboard drainage basins. A similar partitioning of the B. gilchristii populations was observed among the more northerly drainage basins only. These associations suggest that the ecologic niche where the sexual reproduction, growth, and dispersal of B. dermatitidis and B. gilchristii occur is intimately linked to freshwater systems. For most populations, sexual reproduction was rare enough to produce significant linkage disequilibrium among loci but frequent enough that mating-type idiomorphic ratios were not skewed from 1:1. Furthermore, the evolutionary divergence of B. dermatitidis and B. gilchristii was estimated at 1.9 MYA during the Pleistocene epoch. We suggest that repeated glaciations during the Pleistocene period and resulting biotic refugia may have provided the impetus for speciation as theorized for other species associated with temperate freshwater systems.

Predictors of undiagnosed herpes simplex virus type 2 seropositivity among persons attending an HIV care clinic.

We evaluated herpes simplex virus type 2 (HSV-2) sero- positivity in an HIV clinic- based population with CD4 lymphocytes counts ≥250 cells/μL and no previous knowledge of HSV-2 infection by history of serology. We demonstrate that although the seroprevalence of HSV-2 is higher in this HIV-infected population, predictors of HSV-2 seropositivity are similar to those in the general population.

Epidemiology and Geographic Distribution of Blastomycosis, Histoplasmosis, and Coccidioidomycosis, Ontario, Canada, 1990–2015

Elevated incidence of blastomycosis in Ontario calls for diagnostic vigilance.